HTScan® Profiling Kit (Tyrosine Kinase Set I) (#7405) Datasheet with Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

UniProt ID:

#P54753, #P00533, #P22607, #P08581, #P09619, #P08069, #P36888, #P35968

Entrez-Gene Id:

2049, 1956, 2261, 4233, 5159, 3480, 2322, 3791

Product Information

Storage

Antibodies are supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Do not aliquot the antibodies. Store at -20°C. Peptides are supplied at 6 µM in 0.001% DMSO or a DMSO and carbonate buffer solution (15 mM Na2C03, 35 mM NaHC03, 0.05% NaN3). Store at -20°C. Enzymes are supplied in 50 mM Tris-HCL (pH 8.0), 100 mM NaCl, 5 mM DTT, 15 mM reduced glutathione and 20% glycerol. Store at -80°C.
Keep enzmes on ice during use.
Avoid repeated freeze-thaw cycles.

Specificity / Sensitivity

The purity of each RTK was characterized using SDS-PAGE, and assay conditions for phosphorylation of the included substrates were optimalized using DELFIA^® (for details, visit www.cellsignal.com/drug2.AS) (DELFIA^® is a registered trademark of PerkinElmer, Inc.).

Source / Purification

Each GST-Kinase fusion protein in this kit was produced using a baculovirus-expressed cDNA fragment with the target human tyrosine kinase domain amino-terminally fused to a GST-HIS6-Thrombin cleavage site. Proteins were then purified by one-step affinity chromatography using glutathione-agarose.
The detection antibody, Phospho-Tyrosine Mouse mAb (P-Tyr-100) #9411, was corresponding to mice immunized with phospho-tyrosine-containing peptides .

Product Description

The kit provides a means of characterizing the effect of a compound or panel of compounds on the included active human receptor tyrosine kinases.
The tyrosine kinases in the kit are supplied as GST fusion proteins. Biotinylated substrate peptides validated for each kinase are supplied at 6µM, and a phosphotyrosine monoclonal antibody for detection of the phosphorylated form of the substrate peptide is also included in the kit. The kit enables rapid characterization of the effect of a single compound at multiple concentrations or a panel of compounds toward the following RTKs: PDGFR-beta, EGFR, FGFR-3, VEGFR-2, FLT3, IGF1R, MET and EPHB3, each representing an RTK subgroup.

Background

Tyrosine kinases have been validated as targets for therapeutic intervention in many disease areas. The human genome codes for nearly one hundred tyrosine kinases, which can be categorized into receptor tyrosine kinases (RTKs) and cytosolic tyrosine kinase. RTKs are currently a major focus for therapeutic drug target screening in the pharmaceutical industry. HTScan¬Profiling Kit (Tyrosine Kinase Set 1) from Cell Signaling Technology (CST) is an antibody-based kinase activity assay kit.

Each kinase is paired with a validated biotynilated substrate peptide, provided in 96-well format. Optimal assay conditions have been identified for each substrate peptide using a phosphotyrosine-specific antibody to achieve signal-to noise ratios over 25.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

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Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.